Rutaceae Plants Research Articles

Cytotoxic and anti-inflammatory activity of some Thai medicinal plants

In order to search for new anti-inflammatory agent for use as an adjunctive treatment for periodontal diseases, we have studied crude extracts from plants of family Rutaceae, Asteraceae and Maliaceae which have been claimed to possess anti-microbial and anti-inflammatory activities. The cytotoxic effect of these plant crude extracts on human gingival fibroblasts (HGFs) and monocytes was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Then, a non-cytotoxic dose was chosen for each plant extract and used to determine the anti-inflammatory activity. The enzyme linked to immunosorbent assay was used to assay tumor necrosis factor-alpha (TNF-a) and interleukin-1beta (IL-1b) released from cells treated with bacterial lipopolysaccharides (LPS). The results showed that the half maximal inhibitory concentration (IC50) of plant extracts for HGFs were in the range of 308.12 to 1168.66 µg/ml while the IC50 for macrophages were in the range of 54.6 to 90.92 µg/ml. Stem bark of Cratoxylum formosumdemonstrated a comparable inhibitory activity to dexamethasone (DM) on TNF-a production by both cells, but showed detectable reduction of IL-1b only in macrophages. Other plant extracts showed no significant difference for reduction of inflammatory cytokines from macrophages. Moreover, leaf and branch of Murraya paniculata showed greater inhibitory activity than DM on the production of TNF-a from stimulated HGFs. Therefore, these plant extracts may be good candidates for further drug development for adjuctive treatment of periodontal diseases. Key words: Cytotoxic effect, anti-inflammatory activity, Rutaceae, Asteraceae, Maliaceae.

MARKET ASSESSMENT OF HERBAL SUPPLEMENTS IN ROMANIA FOR A PERIOD OF 10 YEARS USING A HOLISTIC APPROACH

s. PHYTOPHARM 2012 M 102 Obzory po kliniceskoj farmacologii i lekarstvennoj terapii [Reviews of clinical pharmacology and drug therapy] ТОМ 10/2012/2 MARKET ASSESSMENT OF HERBAL SUPPLEMENTS IN ROMANIA FOR A PERIOD OF 10 YEARS USING A HOLISTIC APPROACH © Stoia Mihaela , Oancea Simona , Coman Diana 3 Faculty of Medicine, Lucian Blaga University of Sibiu, Romania Department of Agricultural Sciences, Food Industry and Environmental Protection, Lucian Blaga University of Sibiu, Romania Faculty of Engineering, Lucian Blaga University of Sibiu, Romania Almost nonexistent in the early ‘90s, herbal supplements market gained an important dimension to Romanians, with the growing awareness and the amounts that people are willing to pay for improving their health. Since this segment is difficult to monitor by market research companies, we propose in this paper a pleading in favor of increasing consumption of therapeutic and aromatic plant products, based on a holistic national approach. following a 10-year survey (period 2001–2010), we analyzed some specific statistical parameters, such as: top and value sales of dietary supplements in Romania; the average consumption of herbal supplements per capita; share of organic crop areas in the used agricultural area; the turnover of indigenous industry in harnessing the potential of ethnopharmacology; educational attainment by age group; death rates of chronic diseases; life expectancy at age 65; existence of data-bases accessible via internet as a useful tool for information and international promotion of Romanian medicinal plants (420 species); surveys on the treatment of diseases with natural products versus synthetic drugs. in conclusion, if 10 years ago herbal supplements market was almost insignificant, currently, the population shows flexibility in choosing products that are based on active natural substances, especially the social categories with high education and increased access to information. Even sales have tripled in the period 2008–2010, however remains an annual average consumption per capita of 9.5 euros, while in countries like Poland, Czech Republic or Slovakia values vary between 12–15 euros/capita, rising in Western Europe up to c25. Approximately 85 % of the raw material used by Romanian manufacturers comes from native flora. On education, health and life expectancy, these parameters improved over the studied period, the largest increase (25 %) occurring in the educational attainment of the 55–64 age group, which is also the age of onset of many chronic and degenerative diseases. Acknowledgement: This work was supported by a grant of the Romanian National Authority for Scientific Research, CNCS — UEfiSCdi, project number PN-ii–id-PCE-2011–3-0474. ANTI–INFLAMMATION EFFECT OF THAI HERBAL EXTRACTS FOR POTENTIAL ORAL USE © Rodanant Pirasut, Surarit Rudee, Sricha Rajporn, Korsuwanwong Suwanna Faculty of Dentistry, Mahidol University, Bangkok, Thailand This work describes studies on the crude extracts from plants of family Rutaceae, Asteraceae and Maliaceae, which have been claimed to possess anti-microbial and anti-inflammatory activity. Cytotoxicity of these extracts on human gingival fibroblasts (hGf) (CRl 2014) and monocytes (U937) were tested using the 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. in addition, TNf-α, il-1β and il-6 released from hGfs and monocytes treated with bacterial lipopolysaccharides (lPS) were determined using the enzyme-linked immunosorbent assay (EliSA). The results showed that the iC 50 of plant extracts for hGfs were in the range of 308.1 to 1168.7 mg/ml, while the iC 50 for macrophages were in the range of 54.6 to 90.9 mg/ml. Stem bark of Cratoxylum formosum demonstrated a comparable inhibitory activity to standard medication, dexamethasone, on TNf-α released from both cells, but showed detectable reduction of il-1β only in macrophages. Other plant extracts did not show any significant reduction in inflammatory cytokines from macrophages. Moreover, leaf and branch of Murraya paniculata showed greater inhibitory activity than dexamethasone on the production of TNf-α from stimulated hGfs. Therefore, these plant extracts may be good candidates for further drug development, especially for use in treatment of periodontal diseases. This work is supported by the grant from the faculty of dentistry, Mahidol University.

Himalayan Plant Species as Pesticidal Agents

The control of diseases and insect, pests for securing world is important issue for us. A significant way to find a better treatment for crop is the search for insecticidal, pesticidal and antimicrobial constituents. From many years research in biology and biochemistry has made it possible a complete new approach for the protection of plants. As part of our Botanical pesticide research project, numerous compounds from Himalayan plant species of Uttarakhand, India with potential antifeedant activity have been identified. The plants selected for complete investigations were Boenninghausenia albiflora, Skimmia anquetillia, Glycosmis arborea, Vitex negundo, Premna barbata, Callicarpa arborea and Clerodendron indicum and the pests selected were the Plecoptera reflexa, the popular defoliator Clostera cupreta, bamboo leaf roller Crypsiptya coclesalis and polyphagous pest Spodoptera litura. The extracts and pure compounds were also tested against plant pathogens, Agrobacterium tumifaciens, Pseudomonas syringae and Pactobacterium carotovorum for antibacterial activity. In all tested plants B. albiflora and S. anquetillia showed potent insecticidal and antibacterial properties. In addition all investigated plants of Rutaceae and Verbenaceae family were found to be active against S. litura pest. Keywords: Botanical pesticides, Boenninghausenia albiflora, Callicarpa arborea, Clerodendron indicum, Glycosmis arborea, Premna barbata, Skimmia anquetillia, Vitex negundo, Mentha Piperita, Cydia Pomonella, Canavanine, Tridecanone

Response of Diaphorina citri Kuwayama (Hemiptera: Psyllidae) to Volatiles Emitted from Leaves of Two Rutaceous Plants

Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is an important worldwide pest of citrus, because of its ability to vector the three phloem-restricted bacteria in the genus Candidatus Liberibacter, that cause citrus greening or huanglungbing. Studies were conducted to examine the effects of flush growth stage and volatile compounds emitted by two rutaceae plants on oviposition and feeding site selection of D. citri. Three methods including choice, no choice and Y tube olfactometer assays were conducted using plants with only newly expanded leaves (4-6 days old), plants with all leaves completely hardened (14-21 days old) and plants without leaves (14-21 days old). The results showed that significant number of adult D. citri were attracted to plants with newly expanded leaves for oviposition and feeding in both choice and no choice assay, and when the two plants were assayed together in Y tube, significant number of the adults select Y tube arm with Citrus suhuiensis leaves and similarly in choice assay more adults select C. suhuiensis for oviposition and feeding than Murraya paniculata. The results gave an insight into response of D. citri to different plant forms and volatiles emitted by the plant which is useful in monitoring and management of the pest.

High divergence levels of Panonychus citri populations on Rutaceae and Oleaceae as indicated by internal transcribed spacer 1 (ITS1) sequences

The citrus red mite, Panonychus citri (McGregor) (Acari: Tetranychidae), has a worldwide distribution and is the most important citrus pest in many countries, including Japan, Spain and China. Apart from citrus, P. citri also feeds on other woody plants such as pear, peach and holly. To characterize the population genetic structure of citrus red mite in association with their host plants, the gene flow (Nm) and genetic differentiation among sympatric populations of P. citri (F ST estimates) on different host plants were estimated using the ribosomal internal transcribed spacer 1 (ITS1) sequence. The results indicated that there were high levels of gene flow (Nm = infinite) among the populations from the same host genera and the same host family. Populations of P. citri displayed genetic differentiation and low levels of gene flow (Nm < 1) between the populations collected from Rutaceae and Oleaceae host plants. While the haplotype network presented a parallel pattern, there was no one central common haplotype. Pairwise F ST indicated that in China, P. citri has differentiated into two groups, one of which was associated with sweet olive and the other with other host plants. The result of cytochrome oxidase subunit I analysis also supported this result, suggesting the possibility of sympatric host race formation in this species.

New 2-Aryl-1,4-naphthoquinone-1-oxime Methyl Ether Compound Induces Microtubule Depolymerization and Subsequent Apoptosis

In this study, we describe the antitumor activity of QO-1, one of the new 2-aryl-1,4-naphthoquinone-1-oxime methyl ether derivatives. QO-1 is a derivative of macarpine, a natural occurring product from Rutaceae plant. It could potently inhibit cell growth when tested on 19 cancer cell lines. To investigate its mechanism, two cell lines (HeLa and MCF-7) sensitive to QO-1 were selected. Based on flow cytometry, it was found to induce G2/M-phase arrest. Moreover, it could cause microtubule depolymerization both in vitro and in vivo. On the other hand, QO-1 activated spindle assembly checkpoint (SAC) proteins. Expression of Bub1, one of the SAC, was gradually increased, reaching a peak after 16 – 20 h, and then gradually decreased. Instead, QO-1 increased the sub-G1 population, which suggested a cell death population. Actually, expression of Bcl-2 family proteins and activation of caspase-3/7 were evidences of apoptosis. Consistent with these results, cells with DNA fragmentation and multinucleated cells were increased time-dependently after QO-1 exposure. In conclusion, QO-1 has promising antitumor effects via microtubule depolymerization.

Larval Feeding Stimulants for a Rutaceae-Feeding Swallowtail Butterfly, Papilio xuthus L. in Citrus unshiu Leaves

Larvae of a swallowtail butterfly, Papilio xuthus L., feed exclusively on plants of the family Rutaceae, including various Citrus crops. Larvae were strongly stimulated to feed on paper strips impregnated with ethanolic extracts of host-plant leaves. Stimulation of feeding on extracts of Citrus unshiu leaves required a mixture of chemicals including sugars (D: -glucose, D: -fructose, and D: -sucrose), a betaine [(-)-stachydrine], a cyclic peptide (citrusin I), a polymethoxyflavone (isosinensetin), and the lipids 1-linolenoylglycerol, 1-linoleoylglycerol, 1-octadecenoylglycerol, 1-stearoylglycerol, and 1,2-dilinolenoyl-3-galactosyl-sn-glycerol. When these compounds were assayed individually, few larvae consumed test strips. However, larvae readily chewed the test strips treated with a mixture of all compounds, indicating that host recognition by P. xuthus larvae is mediated by a specific combination of both primary and secondary substances. Comparison of 11 stimulant components with 10 compounds from C. unshiu leaves previously reported as stimulant components for oviposition by P. xuthus adult females revealed only one compound, stachydrine, as an ingredient in common. While the larval feeding-stimulant mixture is dominated by nutrients and other compounds of general significance for primary metabolism, the component oviposition stimulants are mostly secondary substances, including flavonoid glycosides, protoalkaloids, a cyclitol, and a betaine, that have restricted distributions in plants. Reliance by adult females on unique profiles of secondary compounds presumably reflects the need to locate and recognize specific host-plant species within a diverse flora. Since the initial host choice for the larvae is made typically by the ovipositing female, however, unique secondary compounds may be less important for larval feeding than are compounds useful for indicating food and microhabitat quality once on the host plant.

Inhibitory Effects of Furanocoumarin Derivatives in Kampo Extract Medicines on P-Glycoprotein at the Blood-Brain Barrier

Furanocoumarin derivatives, known as components of grapefruit juice, showing inhibitory effects against P-glycoprotein (P-gp) in the intestine are also contained in the plants of rutaceae and umbelliferae families, which are used as components of Kampo extract medicines. In this study, we investigated the inhibitory effects of byakangelicol and rivulobirin A, known as furanocoumarins showing P-gp inhibitory effect using Caco-2 monolayer, against P-gp at the blood-brain barrier (BBB) under both in vitro and in vivo conditions. First we studied the membrane permeability of furanocoumarins to clarify whether they can be absorbed from the intestine. Both furanocoumarins showed high permeability through the Caco-2 monolayer, suggesting that they can easily reach the systemic circulation after oral administration. Then, we evaluated the effect of these furanocoumarins on the uptake of calcein acetoxymethyl ester (calcein-AM), a P-gp substrate, into bovine brain microvascular endothelial cells (BBMEC). Both furanocoumarins significantly increased the uptake amount of calcein-AM into BBMEC by the inhibition of P-gp at the BBB in vitro. Next we also investigated the P-gp inhibitory effect of these furanocoumarins at the rat BBB in vivo using verapamil as a P-gp substrate. Both furanocoumarins increased the B/P ratio of verapamil compared to the control, even under in vivo conditions; however, the extent of the inhibitory effect was much lower than in vitro condition. In conclusion, byakangelicol and rivulobirin A may inhibit P-gp expressed at the BBB even under in vivo conditions. Further studies using Kampo extract medicines under in vivo condition are necessary for safe drug therapy.

Furanocoumarin Derivatives in Kampo Extract Medicines Inhibit Cytochrome P450 3A4 and P-Glycoprotein

Furanocoumarins in grapefruit are known to show inhibitory effects against P-glycoprotein (P-gp) and CYP3A4 in intestinal epithelial cells; however, furanocoumarin derivatives are widely contained in the plants of Rutaceae and Umbelliferae families, which are used as components of Kampo extract medicines. In this study, we investigated the inhibitory effects of 12 furanocoumarins extracted from plants in the Umbelliferae family against P-gp and CYP3A4 activity. Furthermore, we studied their inhibitory effect on P-gp when furanocoumarins are used as Kampo extract medicine rather than as an isolated single compound. From screening of the CYP3A4 inhibitory effect, notopterol and rivulobirin A, the only dimer types of furanocoumarin, were found to be potent inhibitors of CYP3A4. On the other hand, byakangelicol and rivulobirin A showed strong P-gp inhibition from the screening of P-gp inhibitor evaluated by quinidine permeation through the Caco-2 monolayer; however, the chemical structural relationship of furanocoumarins between P-gp and CYP3A4 inhibitory effects could not be obtained. We also investigated the effect of these furanocoumarins on the transport of digoxin through the Caco-2 monolayer. The inhibitory effect of rivulobirin A was more potent than that of byakangelicol. Application of either Senkyu-cha-cho-san or Sokei-kakketsu-to, which are composed of herbal remedies in the Umbelliferae group, significantly decreased the efflux ratio of digoxin. In conclusion, it was found that some furanocoumarins extracted from the plants in the Umbelliferae family strongly inhibited P-gp and CYP3A4. Kampo extract medicines containing herbal remedies belonging to the Umbelliferae family may cause a drug-drug interaction with P-gp or a CYP3A4 substrate drug.

Lignans from the Bark of Zanthoxylum planispinum

AbstractA new dimeric lignan, named biplanispine A (1) which represents the first example of a dimeric 2,6‐diaryl‐3,7‐dioxabicyclo[3.3.0]octane skeleton from a Rutaceae plant, together with 13 known lignans 2–14, was isolated from the bark of Zanthoxylum planispinum. Their structures were elucidated by spectroscopic methods, especially 2D‐NMR techniques. All isolated compounds were evaluated for their antibacterial activities by the disc diffusion method. Compounds 7 and 9 showed interesting antibacterial activities at 200 μg/disc; they were as effective as kanamycin sulfate at 10 μg/disc against Staphylococcus aureus and Bacillus subtilis.

Micromorphological investigation on the leaves of the rock samphire (Crithmum maritimum L.): Occurrence of hesperidin and diosmin crystals

A micromorphological study was carried out on Crithmum maritimum L. (rock samphire), a halophyte belonging to the Apiaceae and used in folk medicine. The study led to the finding of relevant amounts of two closely related flavonoids, namely diosmin and hesperidin, which attract increasing interest for their biological properties. Anatomical investigations of leaf tissue showed the presence of needle‐shaped crystals forming plumose clusters, mainly located close to, or within, vascular bundles. Scanning electron microscopy (SEM) analysis provided a more detailed morphological characterization of crystals, while the absence of mineral elements in their composition was assessed by SEM‐EDX. High performance liquid chromatography (HPLC) analysis of leaf tissue was conducted to detect hesperidin and diosmin and to quantify their amounts. SEM analysis of two Rutaceae plants rich in hesperidin and diosmin, i.e., Citrus limon and Barosma betulina, revealed the same plumose crystals found in C. maritimum, while in these plants the two flavonoids were also quantified by HPLC. The data clearly indicate the occurrence of hesperidin and diosmin crystals in C. maritimum leaves, thus providing a scientific basis for an exploitation of this plant in saline agriculture, as a crop or source of bioactive phytocompounds.

Screening of Trypanosoma cruzi glycosomal glyceraldehyde-3-phosphate dehydrogenase enzyme inhibitors

The inhibitory activity of crude extracts of Meliaceae and Rutaceae plants on glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) enzyme from Trypanosoma cruzi was evaluated at 100 μg/mL. Forty-six extracts were tested and fifteen of them showed significant inhibitory activity (IA % > 50). The majority of the assayed extracts of Meliaceae plants (Cedrela fissilis, Cipadessa fruticosa and Trichilia ramalhoi) showed high ability to inhibit the enzymatic activity. The fractionation of the hexane extract from branches of C. fruticosa led to the isolation of three flavonoids: flavone, 7-methoxyflavone and 3',4',5',5,7-pentamethoxyflavone. The two last compounds showed high ability to inhibit the gGAPDH activity. Therefore, the assayed Meliaceae species could be considered as a promising source of lead compounds against Chagas' disease.

The Rutaceae Family - Order Sapindales

A thorough description is given of the Rutaceae plant family. A wide variety of sources were used including old and new masters in homeopathy and works on pharmacognosy, phytochemistry, mythology, symbolism, phylogeny and plant systematics.

Solar Simulator-Induced Phototoxicity of the Furoquinoline Alkaloid Dictamnine Compared to 8-Methoxypsoralen and 5-Methoxypsoralen

Dictamnine, a furoquinoline alkaloid of the Rutaceae plant family, has been shown to be mutagenic and phototoxic in bacteria and yeasts. Here, we have investigated the phototoxic effect of dictamnine in human Jurkat T cells and HaCaT keratinocytes. Dictamnine was isolated from the roots of DICTAMNUS ALBA L. and was photoactivated with solar simulated radiation, delivered from a 1000-W xenon arc lamp with a maximal output between 300 - 800 nm. Dictamnine displayed concentration- and light-dependent phototoxic effects in both cell lines. In comparison to the structurally related furocoumarins 5-methoxypsoralen and 8-methoxypsoralen, dictamnine was less phototoxic. Nevertheless, it may play a major role in the elicitation of phytophotodermatitis because of its abundance in plants of the Rutaceae family.

奄美諸島におけるカンキツグリーニング病の発生調査（第１報 奄美諸島における分布の特徴）

Citrus Huanglongbing (syn. greening disease) caused by 'Candidatus Liberibacter asiaticus' were surveyed on the Amami islands from 2002 to 2006. A total of 212, 587 Rutaceae plants were surveyed at 29, 063 sites on the Amami islands. Samples that showed a greening-like symptom (17, 384 samples) were examined by a polymerase chain reaction (PCR) assay according to the method of Jagoueix et al. (1994). Of these, 1, 041 (0.5%) samples were identified as being infected with 'Candidatus L. asiaticus.' These infected samples were found on the Kikai-jima, Tokuno-shima, Okierabu-jima, and Yoron-tou islands; however, infected samples were not found in Amami-O-shima (Amami main island).

Mechanism-Based Inactivation of Human Liver Microsomal CYP3A4 by Rutaecarpine and Limonin from Evodia Fruit Extract

Evodia fruit (Evodiae Fructus) is used as a herbal medicine prepared from the matured fruit of the Evodia rutaecarpa Bentham or Evodia officinalis Dode, of the Rutaceae plant family. An extract of Evodia fruit in the presence of NADPH was shown to inhibit human liver microsomal erythromycin N-demethylation activity, mediated by cytochrome P450 3A4 (CYP3A4), in a preincubation-time dependent manner. The present study was conducted to identify components of Evodia fruit extract having preincubation-time dependent inhibitory effects on CYP3A4 by analyzing human liver microsomal erythromycin N-demethylation activity. Rutaecarpine, a major component of Evodia fruit, and limonin caused the most dramatic decrease in residual CYP3A4 activity (IC50 before and after 20 min preincubation with: rutaecarpine, >100 microM and 1.4 microM; limonin, 23.5 microM and 1.8 microM, respectively). Furthermore, rutaecarpine and limonin were identified as mechanism-based inhibitors of CYP3A4 from the following observations: 1) The inhibitory effects of rutaecarpine and limonin on CYP3A4 activity were dependent on the preincubation time, 2) The inhibition required NADPH, 3) The inhibition was depressed in the presence of the competitive CYP3A4 inhibitor, ketoconazole, 4) Dialysis resulted in no recovery of CYP3A4 activity. The kinetic parameters for inactivation k(inact) and K(I) were: 0.387 min-1 and 107.7 microM for rutaecarpine, 0.266 min-1 and 23.2 microM for limonin, respectively. These results indicate that rutaecarpine and limonin are mechanism-based inhibitors of CYP3A4.

Photo-activated DNA binding and antimicrobial activities of furoquinoline and pyranoquinolone alkaloids from rutaceae.

To find novel photo-active compounds of potential use in photochemotherapy from higher plants, photo-activated antimicrobial and DNA binding activities of the furoquinolines, skimmianine, kokusaginine, and haplopine, and a pyranoquinolone, flindersine, from two species of Rutaceae plants were investigated. TLC overlay assays against a methichillin-resistant strain of Staphylococcus aureus and Candida albicans were employed to test antimicrobial properties. All of the tested compounds showed photo-activated antimicrobial activity against S. aureus in the order of kokusaginine > haplopine, flindersine > skimmianine. Weaker activity was found for C. albicans. Photo-activated DNA binding activity of these compounds was investigated by a method using restriction enzymes and a specially designed 1.5 kb DNA fragment. Kokusaginine showed inhibition against all of the 16 restriction enzymes. Haplopine showed a similar inhibition pattern but the binding activity against Asc I and Sma I with restriction sequences consisting only of G and C was very weak. Skimmianine showed binding activity against Xba I, BciV I, Sal I, Pst I, Sph I and Hind III, but very weak or no activity was found for the other restriction enzymes. A pyranoquinolone, flindersine, showed no activity against any of the restriction enzymes. Photo-activated DNA binding activity of furoquinolines was therefore in the order of kokusaginine > haplopine > skimmianine, which was the same order as their photo-activated antimicrobial activity against S. aureus. Therefore, it was concluded that DNA is one of the cellular targets for the furoquinolines to exert their biological activities, similar to psoralens. However, because flindersine showed photo-activated antimicrobial activity against S. aureus but did not show photo-activated DNA binding activity, it is clear that there are other cellular target components for this compound to exert photo-toxic activity.

Trypanocidal activity of Meliaceae and Rutaceae plant extracts.

The in vitro trypanocidal activity of 22 extracts and 43 fractions of plants belonging to the families Meliaceae and Rutaceae was evaluated. The extracts from leaves of Conchocarphus heterophyllus and branches of Trichilia ramalhoi were the most active. The trypanocidal activity seems to be increased by fractionation of the extracts. Fractions from C. heterophyllus and Galipea carinata were the most active and a 100% lysis of the parasites was observed for five fractions. From one of them were isolated two flavonoids: flavone and 7-methoxyflavone, which showed weak trypanocidal activity. The results obtained from the extracts and fractions revealed that the order Rutales is a promising source for the search of new drugs for Chagas disease. Phytochemical studies with the other active fractions are underway in order to isolate compounds, which could be associated with observed activities.

Decrease of intestinal P-glycoprotein activity by 2 n-propylquinoline, a new oral treatment for visceral leishmaniasis

Drugs currently available for visceral leishmaniasis treatment are potentially toxic, have to be administered by parenteral route and frequently give rise to drug resistance, due to the involvement of P-glycoproteins (P-gp) in Leishmania. The purpose of this study was to investigate a possible inhibitory effect of 2 n-propylquinoline (2 nPQ) on P-gp activity. 2 nPQ is a new oral anti-leishmanial drug that has demonstrated its efficacy in BALB/c infected mice with Leishmania donovani [Antimicrob. Agents Chemother. 37 (1993) 859]. Rat everted gut sacs and human intestinal Caco-2 cell lines were used to study the effect of 2 nPQ on P-gp activity. Our results demonstrate an inhibitory effect of 2 nPQ on the P-gp activity with two P-gp substrates (rhodamine 123 and digoxin), two P-gp inhibitors (cyclosporin A and verapamil), and in two different species. Alone or associated with other active drugs, 2 nPQ would be very useful to control Leishmania Multi-Drug-Resistance and intestinal P-gp in humans with kala-azar. Index Descriptors and Abbreviations: VL, visceral leishmaniasis; 2 nPQ, 2 n-propylquinoline; P-gp, P-glycoprotein; MDR, Multi-Drug-Resistance; Caco-2 cells, human colon adenocarcinoma cell line; BALB/c, Consanguine mouse; HIV, human immunodeficiency virus; DMEM, Dulbecco’s modified Eagle’s medium; PBS, phosphate-buffered salt Galipea longiflora, Rutaceae plant rat; everted gut sacs; LD 50, lethal dose 50.

Anxiolytic properties of botanical extracts in the chick social separation-stress procedure.

The recent growth in sales of natural products labeled as dietary supplements in the United States has renewed scientific interest in the study of the therapeutic effects of multi-component botanical products. This study sought to determine whether botanical extracts derived from the Rutaceae family, Acori graminei, the Magnoliaceae family, Alchemilla vulgaris and Primula veris, which had previously been identified in bioassays as having potential anxiolytic activity, were active in the chick social separation-stress procedure. Eight-day-old chicks received IP injections of test articles 30 min before being tested in the presence of two social companions or in isolation for a 3-min observation period. Dependent measures were: a) latency to adopt a ventral recumbent posture to index sedation, b) number of vocalizations to index separation-distress and c) a composite pain score (comprised of footlift frequency and footlift duration in response to 50 microl of 0.10% formalin injected into the plantar surface of the foot) to index stress-induced analgesia. Proprietal extracts NPS00033 from the Rutaceae plant family and NPS00039 (Relora) from the Magnoliaceae plant family screened positive in this chick model without causing sedation. These results suggest that botanical extracts Relora and NPS00033 may be useful in modulating anxiety states.